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Related Experiment Videos

Sperm analysis by flow cytometry using the fluorescent thiol labeling agent monobromobimane.

J Seligman1, R Shalgi, Y Oschry

  • 1Department of Human Genetics, Sackler School of Medicine, Tel Aviv University, Israel.

Molecular Reproduction and Development
|July 1, 1991
PubMed
Summary

Monobromobimane (mBBr) labeling and flow cytometry reveal sperm thiol-disulfide dynamics during maturation and species differences. This method can identify sperm subpopulations and assess sperm quality.

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Area of Science:

  • Reproductive biology
  • Biochemistry
  • Analytical chemistry

Background:

  • Sperm thiol-disulfide status is crucial for male fertility.
  • Changes in thiol and disulfide content occur during sperm maturation.
  • Accurate methods are needed to assess sperm thiol-disulfide profiles.

Purpose of the Study:

  • To investigate sperm thiol and disulfide content using monobromobimane (mBBr) and flow cytometry (FCM).
  • To analyze species-specific differences and intra-sample heterogeneity in sperm thiol-disulfide status.
  • To evaluate the potential of mBBr-labeled FCM as a sperm quality assessment tool.

Main Methods:

  • Labeling of intact spermatozoa with monobromobimane (mBBr) to detect thiols and disulfides.
  • Analysis of bimane-labeled sperm from various mammalian species using flow cytometry (FCM).

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  • Examination of labeled spermatozoa using fluorescent microscopy.

    • Main Results:

    • FCM demonstrated sperm thiol oxidation to disulfides during epididymal maturation.
    • Significant inter-species variations in mature sperm thiol content were observed via FCM.
    • Intra-sample heterogeneity in thiol content, disulfide levels, and reduction resistance was detected.

    Conclusions:

    • FCM analysis of mBBr-labeled sperm provides a robust method for studying thiol-disulfide dynamics.
    • The technique allows for the identification of distinct sperm subpopulations within samples.
    • mBBr-labeled FCM shows promise as a valuable test for evaluating sperm quality.